mcdb lab - Name MCDB 1BL/EEMB 2L Winter 2008 The Effect of...

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Name MCDB 1BL/EEMB 2L Winter 2008 The Effect of Different Salt Concentrations on the Production of Amylase in Barley Seeds Introduction We performed this experiment to answer the question: What is the effect of different salt concentrations on the production of amylase in barley seeds? Barley seeds produce amylase in the presence of Gibberellic Acid (GA) which is used to break down starch. This led us to investigate the effects of salt on amylase production because barley seeds will still grow in the presence of numerous hormones. We sought to find a connection between how much amylase is produced and the concentration of salt present. Methods We incubated halved barley seeds with shaking at room temperature in 4 different incubation mediums. The first medium, which was used as a control, contained only 10 μM of GA. The second medium contained 10 μM of GA and 10 mM of NaCl; the third contained 10 μM of GA and 30 mM of NaCl; the fourth contained 10 μM of GA and 50 mM of NaCl. We then added each incubation medium to a Petri dish with 10 imbibed halved barley seeds containing no embryo. After incubating the halved barley seeds for about 48 hours, we measured the amount of amylase produced. 25 μL of each medium was placed in a test tube with 975 μL of H 2 O and 1 mL of starch substrate. After about 3 to 8 minutes, we added 1 mL of iodine and 5 mL of H 2 O; we read the absorbance at 620 nm using the spectrophotometer.
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Malonzo 2 Results We did 3 tests that tested 25 μL of each medium, with various incubation times. Assuming that the medium with 10 μM of GA and 50 mM of NaCl would produce the most amylase, we incubated this for 3 minutes; we incubated the control for 5.5 minutes (Table 1).
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